19 research outputs found
Imaging cell lineage with a synthetic digital recording system
- Author
- Publication venue
- 'American Association for the Advancement of Science (AAAS)'
- Publication date
- 25/02/2020
- Field of study
Cell lineage plays a pivotal role in cell fate determination. Chow et al. demonstrate the use of an integrase-based synthetic barcode system called intMEMOIR, which uses the serine integrase Bxb1 to perform irreversible nucleotide edits. Inducible editing either deletes or inverts its target region, thus encoding information in three-state memory elements, or trits, and avoiding undesired recombination events. Using intMEMOIR combined with single-molecule fluorescence in situ hybridization, the authors were able to identify clonal structures as well as gene expression patterns in the fly brain, enabling both clonal analysis and expression profiling with intact spatial information. The ability to visualize cell lineage relationships directly within their native tissue context provides insights into development and disease
Imaging cell lineage with a synthetic digital recording system
- Author
- Publication venue
- Publication date
- 25/02/2020
- Field of study
Multicellular development depends on the differentiation of cells into specific fates with precise spatial organization. Lineage history plays a pivotal role in cell fate decisions, but is inaccessible in most contexts. Engineering cells to actively record lineage information in a format readable in situ would provide a spatially resolved view of lineage in diverse developmental processes. Here, we introduce a serine integrase-based recording system that allows in situ readout, and demonstrate its ability to reconstruct lineage relationships in cultured stem cells and flies. The system, termed intMEMOIR, employs an array of independent three-state genetic memory elements that can recombine stochastically and irreversibly, allowing up to 59,049 distinct digital states. intMEMOIR accurately reconstructed lineage trees in stem cells and enabled simultaneous analysis of single cell clonal history, spatial position, and gene expression in Drosophila brain sections. These results establish a foundation for microscopy-readable clonal analysis and recording in diverse systems
Synthetic recording and in situ readout of lineage information in single cells
- Author
- Publication venue
- Nature Publishing Group
- Publication date
- 05/01/2017
- Field of study
Reconstructing the lineage relationships and dynamic event histories of individual cells within their native spatial context is a long-standing challenge in biology. Many biological processes of interest occur in optically opaque or physically inaccessible contexts, necessitating approaches other than direct imaging. Here, we describe a new synthetic system that enables cells to record lineage information and event histories in the genome in a format that can be subsequently read out in single cells in situ. This system, termed Memory by Engineered Mutagenesis with Optical In situ Readout (MEMOIR), is based on a set of barcoded recording elements termed scratchpads. The state of a given scratchpad can be irreversibly altered by Cas9-based targeted mutagenesis, and read out in single cells through multiplexed single-molecule RNA fluorescence hybridization (smFISH). To demonstrate a proof of principle of MEMOIR, we engineered mouse embryonic stem (ES) cells to contain multiple scratchpads and other recording components. In these cells, scratchpads were altered in a progressive and stochastic fashion as cells proliferated. Analysis of the final states of scratchpads in single cells in situ enabled reconstruction of the lineage trees of cell colonies. Combining analysis of endogenous gene expression with lineage reconstruction in the same cells further allowed inference of the dynamic rates at which ES cells switch between two gene expression states. Finally, using simulations, we showed how parallel MEMOIR systems operating in the same cell can enable recording and readout of dynamic cellular event histories. MEMOIR thus provides a versatile platform for information recording and in situ, single cell readout across diverse biological systems
Synthetic recording and in situ readout of lineage information in single cells
- Author
- A McKenna
- A Raj
- A Ventura
- A Wasserstrom
- CC Friedel
- D Frumkin
- DF Robinson
- DL van den Berg
- DR Larson
- E Lubeck
- E Lubeck
- F Farzadfard
- GD Evrony
- J Bonnet
- James M. Linton
- JH Camin
- JH Chung
- Joonhyuk Choi
- K Uranishi
- Ke-Huan K. Chow
- Kirsten L. Frieda
- L Cong
- L Nissim
- LA Banaszynski
- Long Cai
- LV Sharova
- M Jinek
- M Molenaar
- M Percharde
- MA Lodato
- Mark W. Budde
- MB Clark
- Michael B. Elowitz
- N Chapal-Ilani
- N Saitou
- P Mali
- RM Kumar
- RR Sokal
- S Behjati
- S Ding
- S Hormoz
- S Hormoz
- S Shah
- Sahand Hormoz
- SD Perli
- SJ Salipante
- SL Shipman
- V Hsiao
- Y Gao
- Zakary S. Singer
- ZS Singer
- Publication venue
- Nature Publishing Group
- Publication date
- 05/01/2017
- Field of study
Reconstructing the lineage relationships and dynamic event histories of individual cells within their native spatial context is a long-standing challenge in biology. Many biological processes of interest occur in optically opaque or physically inaccessible contexts, necessitating approaches other than direct imaging. Here, we describe a new synthetic system that enables cells to record lineage information and event histories in the genome in a format that can be subsequently read out in single cells in situ. This system, termed Memory by Engineered Mutagenesis with Optical In situ Readout (MEMOIR), is based on a set of barcoded recording elements termed scratchpads. The state of a given scratchpad can be irreversibly altered by Cas9-based targeted mutagenesis, and read out in single cells through multiplexed single-molecule RNA fluorescence hybridization (smFISH). To demonstrate a proof of principle of MEMOIR, we engineered mouse embryonic stem (ES) cells to contain multiple scratchpads and other recording components. In these cells, scratchpads were altered in a progressive and stochastic fashion as cells proliferated. Analysis of the final states of scratchpads in single cells in situ enabled reconstruction of the lineage trees of cell colonies. Combining analysis of endogenous gene expression with lineage reconstruction in the same cells further allowed inference of the dynamic rates at which ES cells switch between two gene expression states. Finally, using simulations, we showed how parallel MEMOIR systems operating in the same cell can enable recording and readout of dynamic cellular event histories. MEMOIR thus provides a versatile platform for information recording and in situ, single cell readout across diverse biological systems
Synthetic recording and in situ readout of lineage information in single cells
- Author
- A McKenna
- A Raj
- A Ventura
- A Wasserstrom
- CC Friedel
- D Frumkin
- DF Robinson
- DL van den Berg
- DR Larson
- E Lubeck
- E Lubeck
- F Farzadfard
- GD Evrony
- J Bonnet
- James M. Linton
- JH Camin
- JH Chung
- Joonhyuk Choi
- K Uranishi
- Ke-Huan K. Chow
- Kirsten L. Frieda
- L Cong
- L Nissim
- LA Banaszynski
- Long Cai
- LV Sharova
- M Jinek
- M Molenaar
- M Percharde
- MA Lodato
- Mark W. Budde
- MB Clark
- Michael B. Elowitz
- N Chapal-Ilani
- N Saitou
- P Mali
- RM Kumar
- RR Sokal
- S Behjati
- S Ding
- S Hormoz
- S Hormoz
- S Shah
- Sahand Hormoz
- SD Perli
- SJ Salipante
- SL Shipman
- V Hsiao
- Y Gao
- Zakary S. Singer
- ZS Singer
- Publication venue
- 'Springer Science and Business Media LLC'
- Publication date
- Field of study
Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)
- Author
- A. -G. Wu
- A. C. Ma
- A. K. Au
- Abdel-Aziz A. K.
- Abdelfatah S.
- Abdellatif M.
- Abdoli A.
- Abel S.
- Abeliovich H.
- Abildgaard M. H.
- Abudu Y. P.
- Acevedo-Arozena A.
- Adamopoulos I. E.
- Adeli K.
- Adolph T. E.
- Adornetto A.
- Aflaki E.
- Agam G.
- Agarwal A.
- Aggarwal B. B.
- Agnello M.
- Agostinis P.
- Agrewala J. N.
- Agrotis A.
- Aguilar P. V.
- Ahmad S. T.
- Ahmed Z. M.
- Ahumada-Castro U.
- Aits S.
- Aizawa S.
- Akkoc Y.
- Akoumianaki T.
- Akpinar H. A.
- Al-Abd A. M.
- Al-Akra L.
- Al-Gharaibeh A.
- Alaoui-Jamali M. A.
- Alberti S.
- Alcocer-Gomez E.
- Alessandri C.
- Ali M.
- Alim Al-Bari M. A.
- Aliwaini S.
- Alizadeh J.
- Almacellas E.
- Almasan A.
- Alonso A.
- Alonso G. D.
- Altan-Bonnet N.
- Altieri D. C.
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- Zwerschke W.
- Publication venue
- 'Informa UK Limited'
- Publication date
- 01/01/2021
- Field of study
Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.
- Author
- Abdel-Aziz AK
- Abdelfatah S
- Abdellatif M
- Abdoli A
- Abel S
- Abeliovich H
- Abildgaard MH
- Abudu YP
- Acevedo-Arozena A
- Adamopoulos IE
- Adeli K
- Adolph TE
- Adornetto A
- Aflaki E
- Agam G
- Agarwal A
- Aggarwal BB
- Agnello M
- Agostinis P
- Agrewala JN
- Agrotis A
- Aguilar PV
- Ahmad ST
- Ahmed ZM
- Ahumada-Castro U
- Aits S
- Aizawa S
- Akkoc Y
- Akoumianaki T
- Akpinar HA
- Al-Abd AM
- Al-Akra L
- Al-Gharaibeh A
- Alaoui-Jamali MA
- Alberti S
- Alcocer-GĂłmez E
- Alessandri C
- Ali M
- Alim Al-Bari MA
- Aliwaini S
- Alizadeh J
- Almacellas E
- Almasan A
- Alonso A
- Alonso GD
- Altan-Bonnet N
- Altieri DC
- Alves da Costa C
- Alves S
- Alzaharna MM
- Amadio M
- Amantini C
- Amaral C
- Ambrosio S
- Amer AO
- Ammanathan V
- An Z
- Andersen SU
- Andrabi SA
- Andrade-Silva M
- Andres AM
- Angelini S
- Ann D
- Anozie UC
- Ansari MY
- Antas P
- Antebi A
- AntĂłn Z
- Anwar T
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- Apostolova N
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- Arasaki K
- Araya J
- AraĂșjo WL
- Arden C
- Arguelles S
- Arias E
- Arikkath J
- Arimoto H
- Ariosa AR
- Armstrong-James D
- Arnauné-Pelloquin L
- Aroca A
- Arroyo DS
- Arsov I
- Artero R
- Arévalo M-A
- Asaro DML
- Aschner M
- Ashrafizadeh M
- Ashur-Fabian O
- Atanasov AG
- Au AK
- Auberger P
- Auner HW
- Aurelian L
- Autelli R
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- Aveic S
- Aveleira CA
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- Chen Z-H
- Chen ZJ
- Chen ZS
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- Cheng S-Y
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- Der CJ
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- Zanella I
- Zang QS
- Zanivan S
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- Zaragoza P
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- Zhang KYB
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- Zhang X
- Zhang X-W
- Zhang XD
- Zhang Y
- Zhang Y
- Zhang Y
- Zhang Y
- Zhang Y
- Zhang Y-D
- Zhang Y-Y
- Zhang Z
- Zhang Z
- Zhang Z
- Zhang Z
- Zhang Z
- Zhang Z
- Zhao H
- Zhao L
- Zhao S
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- Zhao X-F
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- Zheng G
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- Zheng X-L
- Zheng Y
- Zheng Z-G
- Zhivotovsky B
- Zhong Q
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- Zhou K
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- Zhou X-J
- Zhou Y
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- Zhou Y
- Zhou Z
- Zhou Z-Y
- Zhu B
- Zhu C
- Zhu G-Q
- Zhu H
- Zhu H
- Zhu H
- Zhu W-G
- Zhu Y
- Zhu Y
- Zhuang H
- Zhuang X
- Zientara-Rytter K
- Zimmermann CM
- Ziviani E
- Zoladek T
- Zong W-X
- Zorov DB
- Zorzano A
- Zou W
- Zou Z
- Zou Z
- Zuryn S
- Zwerschke W
- Ălvarez ĂMC
- Ăvalos Y
- Ănal G
- ĂstĂŒn S
- ÄoliÄ M
- ÄokiÄ J
- Ćœerovnik E
- Publication venue
- 'Informa UK Limited'
- Publication date
- 01/01/2021
- Field of study
In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field
Benchmarked approaches for reconstruction of in vitro cell lineages and in silico models of C. elegans and M. musculus developmental trees.
- Author
- Chow Ke-Huan K
- Chung Verena
- Elowitz Michael B
- Garry Daniel J
- Gong Wuming
- Granados Alejandro A
- Guan Yuanfang
- Han Lu
- Hu Jingyuan
- Jones Matthew G
- Joseph Thomas
- Khodaverdian Alex
- Kwak Il-Youp
- Liu Zhandong
- Mason Michael
- Meyer Pablo
- Peng Jiajie
- Prusokas Alidivinas
- Prusokas Augustinas
- Rao Aditya
- Rao Suhas
- Raz Ofir
- Rennert Phil
- Retkute Renata
- Saipradeep Vangala G
- Salvador-MartĂnez Irepan
- Shang Xuequn
- Shapiro Ehud
- Shendure Jay
- Sivadasan Naveen
- Srinivasan Rajgopal
- Telford Maximilian J
- Wang Robert
- Yosef Nir
- Yu Thomas
- Zhang Hanrui
- Zhang Richard
- Publication venue
- Cell Syst
- Publication date
- 15/06/2021
- Field of study
The recent advent of CRISPR and other molecular tools enabled the reconstruction of cell lineages based on induced DNA mutations and promises to solve the ones of more complex organisms. To date, no lineage reconstruction algorithms have been rigorously examined for their performance and robustness across dataset types and number of cells. To benchmark such methods, we decided to organize a DREAM challenge using in vitro experimental intMEMOIR recordings and in silico data for a C. elegans lineage tree of about 1,000 cells and a Mus musculus tree of 10,000 cells. Some of the 22 approaches submitted had excellent performance, but structural features of the trees prevented optimal reconstructions. Using smaller sub-trees as training sets proved to be a good approach for tuning algorithms to reconstruct larger trees. The simulation and reconstruction methods here generated delineate a potential way forward for solving larger cell lineage trees such as in mouse
Influence of cadmium and salinity in the red alga Pterocladiella capillacea: cell morphology, photosynthetic performance and antioxidant systems
- Author
- A Andosch
- A Latini
- B Lapointe
- BA Macler
- BE Lapointe
- C Gouveia
- CS Lobban
- D Branco
- DH Turpin
- DM Templeton
- EC Oliveira
- EC Schmidt
- EC Schmidt
- F Chow
- F Perreault
- F Scherner
- FAL Toledo
- FL Figueroa
- H Aebi
- I GĂłmez
- I Rocchetta
- J Collén
- J Grzymski
- J Hamilton
- J Ramus
- JA DeBoer
- JG Ryther
- JG Scandalios
- JJ Vergara
- JM Conitz
- JR Andria
- JR Xia
- K Bird
- KE Arnold
- L Brinza
- L Huan
- L Legendre
- L Talarico
- M Kakinuma
- M Kumar
- M Lerdau
- MD Hanisak
- MJ GarcĂa-SĂĄnchez
- MK Daud
- N Mallick
- N Verbruggen
- NS Yokoya
- OH Lowry
- P Parmar
- PK Hepler
- PX Sheng
- R de Paula Martins
- R Santos
- RE Smith
- RJ Ritchie
- RW Santos
- S Katz
- SP Hwang
- T Mock
- T Wernberg
- U Karsten
- U Schreiber
- ZL Bouzon
- Publication venue
- 'Springer Science and Business Media LLC'
- Publication date
- Field of study
Recommended from our members
Expression-Based Cell Lineage Analysis in Drosophila Through a Course-Based Research Experience for Early Undergraduates.
- Author
- Ahmadi Sabrena M
- Ahn Hannah G
- Akhave Samira J
- Allen Donald N
- Alvarez Silverio D
- Ansari Saif
- Asem Bobby
- Asem Neda
- Azarian Nicole A
- Babaesfahani Arezou
- Bahrami Ahmad
- Banerjee Abhik Kumar
- Bhamra Manjeet
- Bhargava Ragini
- Bhatia Rakesh
- Bhatia Subir
- Bumacod Nicholas
- Caine Jonathan J
- Caldwell Thomas A
- Calica Nicole A
- Calonico Elise M
- Calvopina Joseph Hargan
- Chan Carman
- Chan Helen H-L
- Chang Albert
- Chang Chiaen
- Chang Daniel
- Chang Jennifer S
- Charania Nauman
- Chen Jasmine Y
- Chen Kevin
- Chen Lu
- Chen Yuyu
- Cheung Derek J
- Cheung Jesse J
- Chew Jessica J
- Chew Nicole B
- Chien Cheng-An Tony
- Chin Alana M
- Chin Chee Jia
- Cho Youngho
- Choi In Young
- Chou Man Ting
- Chow Ke-Huan K
- Chu Carolyn
- Chu Derrick M
- Chu Maggie K
- Chu Virginia
- Chuang Katherine
- Chugh Arunit Singh
- Cubberly Mark R
- Daniel Michael Guillermo
- Datta Sangita
- Dhaliwal Raj
- Dickan Joseph N
- Dinh Jenny
- Dixit Dhaval
- Do Minh-Tu
- Dowling Emmylou
- Evans Cory J
- Feng Melinda
- From Christopher M
- Furukawa Daisuke
- Gaddipati Himaja
- Gandhi Anand D
- Gon Kimberly
- Gurley Kayla GH
- Han Lisa
- He Johnny
- Johnson Elizabeth B
- Kelley Ross G
- Khan Awais M
- Khan Omair
- Kim Hee Jong
- Kuoy Edward
- Lam Hiu
- Lee Noemi E
- Lee Szuyao
- Liang Letty
- Mai Van Thanh
- Moridzadeh Naseem
- Ngo Kathy T
- Nguyen Joseph Duy
- Olson John M
- Ou Kristy
- Padhiar Manish M
- Patananan Alexander N
- Patel Vijay
- Sitz Daniel
- Sumra Kirandeep
- Ta Truc
- Tigranyan Annie
- Tran PhuongThao
- Truong-N Khoa T
- Wang Jay
- Wu Jessie C
- Yeh George
- Yoon Earl
- Yu John Y
- Zaretsky Jesse M
- Publication venue
- eScholarship, University of California
- Publication date
- 01/11/2019
- Field of study
A variety of genetic techniques have been devised to determine cell lineage relationships during tissue development. Some of these systems monitor cell lineages spatially and/or temporally without regard to gene expression by the cells, whereas others correlate gene expression with the lineage under study. The GAL4 Technique for Real-time and Clonal Expression (G-TRACE) system allows for rapid, fluorescent protein-based visualization of both current and past GAL4 expression patterns and is therefore amenable to genome-wide expression-based lineage screens. Here we describe the results from such a screen, performed by undergraduate students of the University of California, Los Angeles (UCLA) Undergraduate Research Consortium for Functional Genomics (URCFG) and high school summer scholars as part of a discovery-based education program. The results of the screen, which reveal novel expression-based lineage patterns within the brain, the imaginal disc epithelia, and the hematopoietic lymph gland, have been compiled into the G-TRACE Expression Database (GED), an online resource for use by the Drosophila research community. The impact of this discovery-based research experience on student learning gains was assessed independently and shown to be greater than that of similar programs conducted elsewhere. Furthermore, students participating in the URCFG showed considerably higher STEM retention rates than UCLA STEM students that did not participate in the URCFG, as well as STEM students nationwide